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. 2023 Jul 24;14:4450. doi: 10.1038/s41467-023-39700-1

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 6 — a–c We optimized the coding sequence of human Neurofilament Light Chain, hNFL (NM_006158.5), for C. elegans expression codon biases (see “Methods”), tagged with mNeonGreen fluorophore and expressed under the mec-7 touch neuron promoter. Scale bar = 2 µm. a Strain is expressing bzEx269[P_mec-7mNG::hNFL] and bzSi34[P_mec-7mSc::IFD-1]. Image is representative of N > 20 ALMs. Graphed as Mander’s overlap coefficient of mScarlet channel over mNeonGreen channel, N = 10 ALMs graphed. Boxes indicate coefficient range; minimum (0.51), maximum (0.97), and mean (0.80) coefficient. b Adult touch neuron expressing bzEx313[P_mec-7mNG::UBQ-2]; bzSi43[P_mec-7mSc::hNFL]. Image is representative of N > 10 ALMs. Mander’s overlap coefficient graphed with boxes indicating coefficient range minimum (0.06), maximum (0.84), and mean (0.57). c Representative adult touch neuron expressing single copy bzSi39[P_mec-7mNG::HttQ74]; bzSi43[P_mec-7mSc::hNFL]. Image is representative of touch neurons that display visible mNG aggregation, N > 10 ALMs. Mander’s overlap coefficient plotted; N = 10 ALMs that display visible mNG aggregation. Graphed as Mander’s overlap coefficient; boxes indicate coefficient range; minimum (0.23), maximum (0.99), and mean (0.73). d hNFL-labeled aggresome-like foci can be extruded from touch neurons in exophers; representative image (N > 10 ALM exophers). Strain expresses bzEx311[P_mec-7mNG::hNFL] with bzIs166[P_mec-4mCherry] in the ifd-2(bz187) background. Image shows ALMR neuron with three exophers. Two exophers are positioned in the middle of the image surrounded by scattered mCherry debris, starry night phenotype (SN) from hypodermal exopher digestion⁶. The third exopher pictured, a mature distant exopher, contains a large mNG::hNFL inclusion, shown in focus and zoomed in. The exposure is altered to show details. Out of N = 34 Ad2 ALM exophers from a strain expressing bzSi48[P_mec-7 mNG::hNFL]; bzIs166[P_mec-4mCherry], 16 exophers included mNG::hNFL puncta. e Touch neuron expressed single copy P_mec-7mNG::hNFL partially rescues the exopher suppression phenotype of the ifd-2(bz187) mutant. We measured Ad2 exophers in ALMR. bzIs166[P_mec-4mCherry] compared to bzIs166[P_mec-4mCherry]; ifd-2(bz187) (****P = 3.9 × 10⁻⁹) and bzIs166[P_mec-4mCherry]; ifd-2(bz187) compared to bzIs166[P_mec-4mCherry]; ifd-2(bz187); bzSi48[P_mec-7mNeonGreen::hNFL] (**P = 0.0046), N = 200 ALMR per strain, CMH test, 4 trials.